Genetic analysis, using P22 challenge phage, of the nitrogen activator protein DNA-binding site in the Klebsiella aerogenes put operon.
The nac gene product is a LysR regulatory protein required for nitrogen regulation of several operons from Klebsiella aerogenes and Escherichia coli. We used P22 challenge phage carrying the put control region from K. aerogenes to identify the nucleotide residues important for nitrogen assimilation control protein (NAC) binding in vivo. Mutations in an asymmetric 30-bp region prevented DNA binding by NAC. Gel retardation experiments confirmed that NAC specifically binds to this sequence in vitro, but NAC does not bind to the corresponding region from the put operon of Salmonella typhimurium, which is not regulated by NAC.
Chen
L M
LM
Department of Microbiology, University of Illinois, Urbana 61801, USA.
Goss
T J
TJ
Bender
R A
RA
Swift
S
S
Maloy
S
S
D026921:Amino Acid Transport Systems, Neutral; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D017100:Bacteriophage P22 / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D004269:DNA, Bacterial; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009876:Operon; D012486:Salmonella typhimurium / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
D026921:Amino Acid Transport Systems, Neutral; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C072712:NAC protein, bacteria; C489526:Nac protein, E coli; C080683:PutA protein, Bacteria; D014157:Transcription Factors; C033082:proline transporter